Melatonin is a neurohormone that ameliorates many health conditions when it is administered as a drug, but its drawbacks are its oral and intravenous fast release. To overcome the limitations associated with melatonin release, cyclodextrin-based nanosponges (CD-based NSs) can be used. Under their attractive properties, CD-based NSs are well-known to provide the sustained release of the drug. Green cyclodextrin (CD)-based molecularly imprinted nanosponges (MIP-NSs) are successfully synthesized by reacting β-Cyclodextrin (β-CD) or Methyl-β Cyclodextrin (M-βCD) with citric acid as a cross-linking agent at a 1:8 molar ratio, and melatonin is introduced as a template molecule. In addition, CD-based non-molecularly imprinted nanosponges (NIP-NSs) are synthesized following the same procedure as MIP-NSs without the presence of melatonin. The resulting polymers are characterized by CHNS-O Elemental, Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric (TGA), Differential Scanning Calorimetry (DSC), Zeta Potential, and High-Performance Liquid Chromatography (HPLC-UV) analyses, etc. The encapsulation efficiencies are 60–90% for MIP-NSs and 20–40% for NIP-NSs, whereas melatonin loading capacities are 1–1.5% for MIP-NSs and 4–7% for NIP-NSs. A better-controlled drug release performance (pH = 7.4) for 24 h is displayed by the in vitro release study of MIP-NSs (30–50% released melatonin) than NIP-NSs (50–70% released melatonin) due to the different associations within the polymeric structure. Furthermore, a computational study, through the static simulations in the gas phase at a Geometry Frequency Non-covalent interactions (GFN2 level), is performed to support the inclusion complex between βCD and melatonin with the automatic energy exploration performed by Conformer-Rotamer Ensemble Sampling Tool (CREST). A total of 58% of the CD/melatonin interactions are dominated by weak forces. CD-based MIP-NSs and CD-based NIP-NSs are mixed with cream formulations for enhancing and sustaining the melatonin delivery into the skin. The efficiency of cream formulations is determined by stability, spreadability, viscosity, and pH. This development of a new skin formulation, based on an imprinting approach, will be of the utmost importance in future research at improving skin permeation through transdermal delivery, associated with narrow therapeutic windows or low bioavailability of drugs with various health benefits. Therefore, it can be classified as a class II category drug according to the Biopharmaceutics Classification System (poorly soluble and highly permeable) 2C6H2 (CH3)OH, 99% (GC)); and Beeswax (bleached) were purchased from Sigma-Aldrich .Sodium hypophosphite monohydrate  and acetonitrile  were purchased from VWR Chemicals BDH .The citric acid , Alfa Aesar, was purchased from ThermoFisher  GmbH.Melatonin  and potassium phosphate monobasic  were purchased from Italia Carlo Erba S. p. a., .Disodium hydrogen phosphate dodecahydrate , 1-hexadecanol ), or cetyl alcohol, were purchased from Fluka Chemie GmbH, Buchs, Switzerland.Ortho-phosphoric acid .−1, dispensed through a 0.22 μm membrane filter, were used throughout the studies.All chemicals were analytical-grade commercial products and were used as received. Deionized water, and water purified by reverse osmosis  with a resistivity above 18.2 MΩcmCyclodextrin-based molecularly imprinted nanosponges (labelled as MIP-NSs) were successfully synthesized following the procedure in ,64 with −1, at a spectral resolution of 4 cm−1 and a sample/background scan number of 8. FTIR spectra were obtained using a versatile Attenuated Total Reflectance mode (FTIR-ATR) sampling accessory with a diamond crystal plate. The FTIR-ATR measurements were performed on dried samples.The synthesized MIP-NSs and NIP-NSs were characterized utilizing Fourier Transform Infrared Spectroscopy (FTIR), using a Perkin Elmer Spectrum Spotlight 100 FTIR spectrophotometer equipped with Spectrum software. The FTIR spectra were gained in the spectral range of 4000–650 cmAll the simulations were carried out with the xTB code (extended Tight Binding) v. 6.4.0 at the semi-empirical level of theory xTB-GFN2 ,66. The 2) flow, and with a heating ramp rate of 10 °C/min. The gas flows applied in the balance and furnace section were 40 mL/min and 60 mL/min. About 10 mg of the sample was weighed in an aluminum pan for analysis.The thermal stability of the synthesized MIP-NSs and NIP-NSs was studied by thermogravimetric analysis (TGA). TGA was carried out using a TA Instrument Thermogravimetric Analyzer (TGA) Q500 from room temperature up to 800 °C, under nitrogen  measurements were carried out using a TA instrument Q200 DSC  on a 2–3 mg sample under a nitrogen flow of 50 mL/min. The empty aluminum pan was used as a reference standard. A heating rate of 10 °C/min was employed in the 20–130 °C temperature range.2O5 as a catalyst were placed in a tin container. The CHNS-O analyses were performed in duplicate. The nitrogen signal enabled the estimation of the melatonin presence in the MIP-NSs.CHNS-O elemental analysis (EA) was carried out to quantify accurate and reproducible carbon, hydrogen, nitrogen, and sulfur content in the synthesized polymers using a CHNS-O Analyser , equipped with the Eager Xperience software (for Windows XP) and a MAS 200R Auto Sampler. 2,5-Bis (5-tert-butyl-2-benzo-oxazol-2-yl) thiophene (BBOT) was used as an external standard for the calibration of the system. About 2.5 mg of each sample and an, approximately, equal quantity of VThe morphology of the synthesized MIP-NSs and NIP-NSs was studied using scanning electron microscopy (SEM). The imaging was conducted with a Zeiss EVO 50  using secondary electrons and a 10 kV accelerating voltage. The samples were placed on the aluminum stub, with the help of a bio-adhesive carbon tape. Before SEM analysis, the polymers were ion-coated with gold using a Baltec SCD 050 sputter coater  for 40 s under vacuum at 60 mA.Zeta potential was measured using a Malvern Zetasizer Nano ZS . Approximately 1 mg of the synthesized sample was dispersed in 1 mL of MilliQ water and shaken for a certain time. During zeta potential analysis at 25 °C, the prepared dispersions were placed in the electrophoretic cell and the particles moved to the electrode that had an opposite charge to them upon the application of an electric field (15 V/cm). Three measurements were carried out, and their average was presented.tm is the weight of the swollen sample at time t, and om is the initial weight of the dry sample [The kinetics of the swelling of synthesized MIP-NSs and NIP-NSs was studied by following their increase in weight when immersed in water. The swelling measurements were performed by immersing 0.5 g of dry powder in deionized water (in 15 mL test tubes) and blending them, in the beginning, using a Vortex Mixer. The test tubes were sealed and maintained at room temperature. After 2 h, the mixtures were centrifuged to obtain a layer of water-bound material and free, unabsorbed water. After removing the supernatant, the residual amount of free water was blotted off using tissue paper, and the weight was recorded. The WAC measurements were performed in duplicate. The water absorption capacity (%WAC) was calculated using the following Equation (1):y sample .The storage stability of melatonin was investigated under respective mediums such as phosphate-buffered solutions of pH = 6.04, 6.82, 7.00, and 7.40.2HPO4·12H2O, and 5.04 g of KH2PO4, 8.58 mL of Na2HPO4·12H2O, and 74.2 mL of KH2PO4); pH = 6.82 ; pH = 7.00 ; and pH = 7.40 . The melatonin solutions were analyzed at different interval times for a long period  with a UV-Vis spectrophotometer.0.5 mg of melatonin was solubilized in 5 mL of each medium. Buffer solutions were prepared as follows: pH = 6.04  for 5 h was also investigated and analyzed by TGA.UV-Vis absorption spectra were obtained with a Hewlett Packard UV-Vis spectrophotometer model 845-3. A 10 mm path-length rectangular quartz cell (Hellma) was employed in the 200–400 nm spectral range. The concentration of the samples was adjusted so that the extinction values did not exceed A = 1.max of melatonin.A value of 223 nm was chosen as the λv:v ratio) EtOH/H2O solution. To this solution, 0.5 g of the NIP-NSs was added, and the dispersion was mildly stirred for 24 h at room temperature. The melatonin-loaded NIP-NSs was recovered through centrifugation and freeze-dried.Melatonin-loaded NIP-NSs was prepared by dissolving 20%  in 10 mL 4:6 , which were placed in a 150-mL beaker. The beaker was directly heated on a hot plate at 75 °C until all the ingredients melted. While this mixture was kept warm, the aqueous phase, made of glycerol, and deionized water was further prepared in a 150-mL beaker and heated on a hot plate at 75 °C.After the aqueous phase reached 75 °C, it was slowly added into the oil phase, under constant stirring, with the aid of an overhead stirrer , until a smooth and uniform paste was reached. The quantities of the ingredients are presented in Due to their dried texture , the MIPFor pre-screening purposes, cream formulations underwent a centrifugation test, using the 4232-D Benchtop + 5228 RPM Tube Rotor  centrifuge at a certain weight and 4000 RPM for the duration of 30, 60, and 90 min to evaluate syneresis, precipitation, or phase separation. At defined time intervals, the formulations were controlled to observe if there was any change .The pH of the cream formulations was measured with the aid of a digital pH meter (Orion model 420 A). A total of 0.10 g of MIP-NSs cream was uniformly dispersed in 25 mL of deionized water and kept for 2 h at room temperature. The pH of dispersions was measured at 25 °C .−1). This enabled the study of the effect of MIP-NSs on cream viscosity. The temperature was maintained at a constant 25 °C by a water bath circulator. The sample was placed between the upper parallel plate and stationary surface with a 1 mm gap size. After sample loading, the gap was closed, and the sample edge was carefully trimmed with a spatula to maintain the proper surface shape during the measurements and avoid errors. Measurements were carried out in steady flow using a protocol with three steps such as (1) Conditioning-sampling, where the sample was equilibrated for 2 min before the experiments to allow the relaxation of the whole structure. A delay of 2 min was applied to measure the initial structure level of the samples before shearing to eliminate any disturbance created by the measuring geometry; (2) Flow-Ramp, increasing the linear shear rate from 0.1 to 100 s−1 in 2 min; and (3) Flow-Peak hold, maintaining a constant shear rate at 100 s−1 over 2 min. To ensure the reproducible state of the samples, the measurements were accomplished in triplicate recording their average. The data obtained were expressed as mean values ± SD.The rheological characterization of prepared creams was evaluated using the Rheometer TA Instruments Discovery HR 1 by following the procedure described in the literature with some modifications ,74,75. TS is the spreadability, M is the weight tied to the upper side, L is the length of the glass slide, and T is the time taken to separate the slide from each other. Spreadability was recorded in g·cm/s.The spreadability of the cream was determined using the parallel plate method following the procedure in the literature with slight modifications . A total2O: ACN  solution. After 20 min, the dispersion was centrifuged at 6000 RPM for 10 min, and the supernatant was recovered as the first extract and then replaced with 5 mL of fresh H2O:ACN (70:30 v:v) solution. The extraction was performed three times. A total of 1.5 mL of each extract was filtered through a membrane filter  and was analyzed with HPLC-UV (λMel = 223 nm). Melatonin was quantified against an external calibration curve with standards  prepared by serial dilution with a mobile phase of a 1000 μg/mL stock solution. The calibration curve (R2 = 0.9999) was used to quantify the melatonin extracts. The measurements were carried out in triplicate. Loading capacity (LC) was defined as the mass of encapsulated melatonin (enm), divided by the total mass of nanosponge (NS) (totm), as presented in Equation (3):The extraction of melatonin was accomplished by stirring 0.1 g of nanosponge in 5 mL of Henm) over its initial mass (inm) during the loading step, as presented in Equation (4) [The encapsulation efficiency (EE) was defined as the mass of encapsulated melatonin (tion (4) :(4)EE , with a pH of 7.4 as the receiving phase.Melatonin-release profiles were evaluated in a phosphate-buffered solution . At fixed intervals , the entire medium (8 mL) from the receptor compartment was withdrawn and replaced with the same volume of fresh medium. Each collected medium was filtered through a membrane filter (0.2 μm polytetrafluoroethylene (PTFE), Millipore, 13 mm Syringe Filter), and the amount of released drug was assessed with HPLC-UV. A standard calibration curve of melatonin at λ = 223 nm was obtained (R2 = 0.9995). The in vitro release tests were performed in triplicate.In vitro release tests of melatonin from cream formulations were conducted on static vertical Franz Diffusion Cells  . The Framax of melatonin). The mobile phase was prepared by mixing water (H2O) and acetonitrile (ACN) , at a pH = 3 , and the elution was isocratic. The total run time was set to 10 min, while the retention time of melatonin was observed at 8 min.High-Performance Liquid Chromatography (HPLC) analysis was carried out at room temperature using a PerkinElmer Brownlee Analytical C18 chromatographic column  connected to a PerkinElmer HPLC system, comprising a Flexar pump working at a flow rate of 1 mL/min and a Flexar UV–vis detector set at 223 nm (λThe polymerization reaction led to a successful synthesis of cyclodextrin-based molecularly imprinted and non-molecularly imprinted nanosponges (MIP-NSs and NIP-NSs). The storage stability of melatonin buffer solutions in pH 6.04, 6.82, 7.00, and 7.40, stored at room temperature, was investigated by a UV-Vis spectrophotometer. It can be observed from The thermal analysis of MIP-NSs and NIP-NSs was analyzed by the TGA a,b. The The changes in the peaks of the TGA curves of β-CD or M-βCD from the MIP-NSs or NIP-NSs were observed due to changes in chemical structure, which led to the formation of polymeric materials.The DSC thermograms a,b prese−1 (O-H stretching vibrations), 2923 cm−1 (C-H stretching vibrations), 1254 cm−1, and 1020 cm−1 (C-O stretch). The cross-linking of citric acid with β-CD or M-βCD was presented by a characteristic band at 1720 cm−1, assigned to the C=O stretching vibrations of the cross-linking group. Whereas the absorption at 1624 cm−1 was due to the carboxylic acid moieties. All the synthesized samples presented comparable spectra with similar bands at characteristic wavenumbers and similar relative intensities, which displayed no interaction of melatonin with MIP-NSs or NIP-NSs .−1, the very broad peak of the hydroxyl group could be found, covering and including the N-H stretching signal , typical of the amide group of melatonin. In the range 3050 to 2900 cm−1, there were the C-H stretching signals, while there was a single, very intense peak representing the stretching of the carbonyl groups of the melatonin amide group at 1730 cm−1. The higher intensity of this peak in the experimental spectrum could be explained by the presence of the carboxyl group of citric acid used as a cross-linking agent to synthesize CD-based nanosponges (CD-based NSs) in an 8:1 molar ratio concerning β-CD. However, a broader shoulder at 1650 cm−1 emerged from the citrate signal of MIP-NSs (green spectrum), in good accordance with the computed ν(CO) amide peak of melatonin. Therefore, it could be concluded that the main peak was the ester CO of citrate, while the peak tailing was due to the CO of melatonin. The digital fingerprint zone showed good superposition among the experimental and computational spectra as well, even for what concerns the relative intensities.A detailed comparison  between Although the computed IR spectrum was based on the single inclusion complex melatonin and β-CD, good agreement between the experimental and computational ones seemed to suggest the proper insertion of the guest molecule (melatonin) inside the cavity, confirming the creation of a multiple series of inclusion complexes, to further lead to the formation of nanosponges (NSs).CD-based NSs are very delicate sponge-like cross-linked structures. The surface structure of cross-linked MIP-NSs and NIP-NSs was further morphologically studied by the SEM technique . SEM imaThe capacity of NIP-NSs to absorb water, as a function of the presence of functional groups in the polymer matrix, was determined. As the network of NIP-NSs bears hydroxyl and carboxylic acid groups, there was a high affinity for water molecules . The watv:v ratio) to affect the solubilization capacity of the melatonin. The ethanol molecules were completely miscible with the water molecules, thus decreasing the dielectric constant. The lower dielectricity of the ethanol/water mixtures led to the solubility of non-soluble drugs compared to pure water [The encapsulation and release behavior of the melatonin loaded into MIP-NSs and NIP-NSs was investigated and quantified by HPLC-UV. Melatonin is poorly water-soluble, but it is soluble in ethanol (20 mg/mL). In this study, the ethanol was mixed with water , in loading capacity, was investigated. With the increase in melatonin quantity in MIP-NSs and NIP-NSs, the loading capacity and encapsulation efficiency significantly increased. CHNS-O elemental analysis confirmed  with the highest encapsulation efficiency (low loading capacity), as previously discussed, presented the lowest drug release percentages. The better-controlled release for MIP-NSs could be explained due to the carboxylic groups, the citric-based MIP-NSs at pH 7.4 (phosphate-buffered solution), which caused the diffusion rate of the buffer into imprinted specific cavities to be slower . The COOAs already observed in the literature , the MIPThe formulations underwent a centrifugation test, and the stability of the final optimized cream was observed at room temperature, using the centrifuge, at a certain weight and 4000 RPM for the duration of 30, 60, and 90 min to evaluate syneresis, precipitation, or phase separation. No changes in the texture of the cream or instability phenomena after 30, 60, and 90 min were observed . Figure Based on this new material proposed for topical melatonin delivery, there is a chance to greatly advance the MIP-NSs development and explore the asset of the transdermal administration over the parenteral and oral routes.v:v, EtOH:H2O). The synthesized NIP-NSs were characterized by water absorption capacity (WAC) of ~250%, a fundamental property that affects drug release rates. Moreover, the stability of NIP-NSs was proven by negative zeta potential values (−24 to −33 mV). No degradation of melatonin  was observed during the storage of MIP-NSs at room temperature, and their exposure to light for several weeks. After melatonin loading, an in vitro release study, using a Franz Diffusion Cell, was performed. It presented the ability of MIP-NSs to control and sustain the release of melatonin, supporting a release mechanism in which the percentage of the drug released from the polymer matrix was dependent on the selective interaction between the imprinted cavities and melatonin. Due to the different associations within the polymeric structure, MIP-NSs presented better-controlled drug release performance (pH = 7.4) for 24 h, with 30–50% more released melatonin than NIP-NSs with 50–70% released melatonin. In this study, a cream formulation (oil/water phase) for the delivery of melatonin was introduced. The applicability of this material, mixed with the formulation, was then evaluated for its stability, spreadability, viscosity, and pH. MIP-NSs, due to the presence of binding sites, seemed to be a very promising polymeric device for the controlled and selective release of melatonin. A non-toxic, biodegradable, and biocompatible imprinting material was introduced, and, despite the high-temperature conditions (140 °C and 100 °C) of its synthesis, the inclusion complex formation (nanosponges/melatonin) was not affected. The same inclusion complex was also supported by the adoption of computational methods through static simulations in the gas phase at the GFN2 level.The green cyclodextrin-based molecularly imprinted nanosponges (MIP-NSs) were successfully synthesized by reacting β-Cyclodextrin (β-CD) or M-β Cyclodextrin (M-β CD), with citric acid as a cross-linking agent at a 1:8 molar ratio in deionized water in the presence of melatonin as a target molecule. In addition, to be compared, β-CD or M-βCD- based non-molecularly imprinted nanosponges (NIP-NSs) were further synthesized and the melatonin, as a target molecule, was loaded (4:6, This study can be used to plan other platforms that can release drugs in a sustained manner. The optimization of the synthesis, characterization, reproducibility, selectivity, recognition, stability, durability, and delivery path of MIP-NSs for a selected template separate to the NIP-NSs is a further research perspective.


The objectives of this study were to evaluate the delamination of convex-shaped metformin HCl (MF) and evogliptin tartrate (EG) multi-layer tablets depending on the pre-compression and main compression pressures and simultaneously correlate these results with those of a surface roughness analysis. Free-flowing MF and EG  granules prepared using the wet granulation method were pre-compressed and subsequently compressed into bilayer and trilayer tablets using a universal testing machine. The compaction force required to break the tablets increased linearly as the main compression pressure increased (30–150 MPa). Conversely, the interfacial strength and compaction breaking force decreased as the pre-compression pressure increased (10–110 MPa). A surface roughness analysis employing a profilometer revealed that the first layer (MF) roughness drastically decreased from 5.89 to 0.51 μm  as the pre-compression pressure increased from 10 to 150 MPa in the bilayer tablet. Accordingly, the decrease in the roughness of the first layer reduced the inter-penetration at the interface, as observed via energy dispersive spectrometer (EDS)-equipped scanning electron microscopy, decreasing the interfacial bonding strength and causing delamination of the MF/EG multi-layer tablets. These findings indicate the significance of roughness control in the actual preparation of multi-layer tablets and the usefulness of profilometer- and EDS-based surface analyses for interpreting the delamination of multi-layer tablets. Multi-layer tablets (MLTs), including bilayer or trilayer tablets, are gaining considerable attention as effective tools for fixed-dose combination (FDC) therapy, with advantages over conventional monolithic tablets ,2. IncomVarious physical and mechanical studies have been conducted to understand the factors that contribute to delamination or cracks in MLTs. Tablet defects are principally associated with interfacial adhesion between adjacent layers and the mechanical integrity of the solid dosage form . MoreoveTherefore, we aimed to evaluate the effect of the compression pressure on the surface roughness of first layer, the compaction force required to break the MLTs, and the interfacial strength, and their correlations in the MLT pharmaceutical product. As a model product, metformin HCl (MF) and evogliptin tartrate (EG) fixed-dose combination tablets, currently prescribed to treat type 2 diabetes mellitus, were employed ,19,20. MIn this study, both EG- and MF-loaded granules were prepared using wet granulation, and their morphology, particle size, density, and flowability were evaluated. The efficiencies (%) of the granulation processes for EG and MF were determined to be 98.7% and 99.8%, respectively. For MF granules, the drug ratio was designed to be approximately 80%, considering the large clinical dose of MF (500–1000 mg/day) . Carbome0.5) of the EG granule was measured to 38.33 μm, with a homogeneous size distribution   . In conty drying . The buly drying . A CI ofwability . Similar2 = 0.9 for both types).The granules were compressed using a universal testing machine  to prepare the MF/EG MLTs. Compression using a universal testing machine provides advantages such as precise control over the compression force and speed, real-time data collection, and understanding of material behavior . The porNext, the porosity and compaction breaking force of the MF/EG bilayer and trilayer tablets depending on the PRE-P were evaluated using a fixed MAIN-P value (150 MPa). As shown in Several methods have been developed to quantify the strength of bilayer tablets, such as the tensile, shear, and diametrical compression; three-point bending; and V-shaped punch-breaking tests ,27,28,292 value of 0.93 for the bilayer tablet , a representative parameter for surface roughness, was employed . As showThe migration of the active ingredient of the first layer into the second layer depending on the PRE-P was further analyzed using EDS-equipped SEM observations. Chang et al.  revealedEDS analysis revealed that in tablets compressed with a low PRE-P , the MF contained in the first layer actively interpenetrated the EG-loaded second layer A. MF, ex® 934P-NF) and mannitol (Pearlitol 100 SD) were obtained from BF-Goodrich  and Roquette , respectively. Low-substituted hydroxypropyl cellulose (L-HPC) and hydroxypropyl cellulose (Klucel LF) were obtained from Shin-Etsu Chemical  and Ashland , respectively. Pregelatinized starch (Starch 1500) and croscarmellose sodium (Ac-Di-Sol) were supplied by Colorcon  and FMC Corp. , respectively. Colloidal silicon dioxide (Aeroperl 300) was purchased from Evonik . Magnesium stearate and red iron oxide (color index number of 77491) were acquired from FACI Asia Pacific  and Univar , respectively.MF (median diameter of 38.33 μm) and EG drug powders (median diameter of 44.74 μm) were obtained from Granules India Limited  and Dong-A ST , respectively. Polyvinylpyrrolidone (PVP K30) was purchased from BASF . High-viscosity grade HPMC2208 (Methocel K100M) and methacrylic acid copolymers (Eudragit S100) were obtained from Dow Chemical  and Evonik , respectively. Carbomer 934P (Carbopolw/w) as the binder solution using an impeller mixer . The solution was sprayed through a nozzle onto the MF drug powder in a fluid bed granulator  to form granules. The parameters of the granulation process were as follows: inlet air temperature of 70 °C, airflow rate of 2/4 m3/h, exhaust temperature of 37–40 °C, and binder solution feeding rate of 500 mL/min. Subsequently, the dried granules were sieved using a 1.2 mm mesh and admixed with HPMC2208 and Carbopol® 934P in a container mixer  at 10 rpm for 10 min. Magnesium stearate, sieved through a 40-mesh sieve, was added to the mixture and lubricated at 10 rpm for 5 min.Both MF- and EG-loaded granules were prepared using wet granulation for a batch of 300,000 tablets . The exaEG-loaded granules were prepared using a high-speed mixer . The drug powder, mannitol, pregelatinized starch, L-HPC, and colloidal silicon dioxide were added to the chamber and mixed for 5 min with an agitator and chopper at speeds of 75 and 1500 rpm, respectively. Then, an aqueous binder solution containing HPC and iron oxide at concentrations of 0.075 and 0.01 mg/mL, respectively, was sprayed into the mixture for 5 min at agitator and chopper speeds of 75 and 1500 rpm, respectively. The wet granules were dried using a fluid bed granulator  for 25 min at an inlet air temperature and an exhaust temperature of 50 and 35 °C, respectively. The dissipated granules were sieved through a 0.8 mm sieve using an oscillator . Croscarmellose sodium was further added to the dried granules as a disintegrant in a container mixer  and mixed at 15 rpm for 10 min. Magnesium stearate, sieved through a 40-mesh sieve, was added to the mixture at 15 rpm for 3 min.0.9–d0.1)/d0.5, where d0.1, d0.5, and d0.9 represent the particle sizes below which 10%, 50%, and 90% of the sample particles lie, respectively [The prepared drug-loaded granules were characterized in terms of drying (LOD), morphology, particle size, density, and flowability (CI and HR). The LOD (%) of the granules (approximately 5 g) was determined using a halogen moisture analyzer  at 105 °C for 15 min until an equilibrium state was reached . For morectively . The BD ectively . GranuleMF/EG-loaded bilayer and trilayer tablets were fabricated by sequential compression of EG- and MF-loaded granules, as depicted in In the case of the trilayer tablets, MF-loaded granules were manually filled into the die and pre-compressed at different PRE-Ps (10–110 MPa) to form the first layer. Then, the EG granules were manually filled in the first layer and were subsequently pre-compressed with the same pressure as the first PRE-P. Finally, the MF-loaded granules were added to the bilayer and subsequently compressed at compression pressures of 30–150 MPa. The thicknesses of the trilayer tablets ranged between 6.56 and 6.75 mm.ε) of MLTs compressed with different PRE-Ps and MAIN-Ps, the true density was measured by measuring the true volume of the sample using a helium Ultrapyc 1220e Automatic Gas Pycnometer . Then, the porosity (ε) was calculated using the following the equation: tρ is the true density, m is the weight of the tablet, and v is the volume of the tablet. The apparent volume of the tablet was calculated using a 3D modeling program .To determine the porosity . The prepared MLTs were placed between two plates and compressed on the tablet at a speed of 1 mm/min, causing fracture or delamination of the MLTs. The peak force obtained from the force–displacement plots was determined as the compaction force required to break the tablets.σ) was calculated as the maximum axial interfacial tensile force (F) divided by the cross-sectional area (A) of the tablet using Equation (2).The interfacial tensile strength of MF/EG-loaded MLTs was determined using a universal testing machine  as previously reported . A newlyThe surface roughness of the first layer following pre-compression was profiled using stylus profilometry . A diamond-tip stylus (radius of 2 μm) was drawn along the first layer of the bilayer tablet with a movement speed of 285 µm/s and a tracking force of 1 mg. The scan size and temperature were set to 2000 µm and 25 °C, respectively. In total, 2100 points were scanned per tablet and the degree of surface roughness was characterized in accordance with International Standards Organization 4287  as folloRp is largest profile peak height and Rv is the largest profile valley depth in one sampling length.The average maximum height of the profile Rz (Equation (3)) is the sum of the maximum profile peak height and the maximum profile valley depth for one sampling length.Rq (Equation (4)), is the root mean square average of the roughness profile ordinates.The root mean square roughness, Ra (Equation (5)) is the arithmetic average of the absolute values of the roughness profile ordinates.l is the scan length of the sample and Z(x) represents the height of the profile at each position x.The average roughness The roughness of the first layer after pre-compression was examined using SEM . The pre-compressed MF layer was mounted onto a copper stub with the surface facing up using double-sided carbon tape . The samples were then coated with a thin layer of Pt. Microphotographs of the coated samples were obtained using scanning electron microscopy at an acceleration voltage of 15 kV.An EDS  equipped with a scanning electron microscope was further employed to track chloride (Cl) originating from MF at the interface and adjacent layers. After the pre-compression and subsequent main compression processes, the interface between the MF and EG layers was cross-sectioned using a razor. Samples were then mounted on a stud using double-sided electrical carbon tape. The energy resolution of the Si drift detector was set to 125 eV.Each experiment was performed at least three times, and the data are presented as the means ± standard deviations (SD). The linearity between the data was analyzed using Origin software . The effects of the PRE-P and the MAIN-P on the interfacial roughness, compaction breaking force, and interfacial strength of convex-shaped EG and MF MLTs were successfully evaluated using a profilometer and EDS-equipped SEM. A roughness analysis using a profilometer revealed that increasing the PRE-P led to a decrease in the surface roughness of the first layer, thus weakening the intermolecular adherence between the adjacent layers and decreasing the interfacial strength required to delaminate the tablets. Accordingly, using EDS-SEM, the inter-penetration at the interface was diminished when the roughness of the first layer was decreased in MF/EG MLTs. Therefore, we suggest that observing the surface roughness of the first layer using a profilometer and EDS-SEM can be a simple and effective tool to understand the delamination phenomenon of MLTs.


The aim of this systematic review and meta-analysis was to evaluate the safety and effectiveness of endoscopic carpal tunnel release (ECTR) using best-evidence synthesis methods.A systematic search of multiple databases was conducted for prospective contemporary studies published between January 2013 and January 2023 with at least 50 ECTR cases. Outcomes included the Quick Disabilities of the Arm, Shoulder, and Hand Questionnaire (Q-DASH) measured on a 0–100 scale, Boston Carpal Tunnel Questionnaire Symptom Severity Scale (BCTQ-SSS) and Functional Status Scale (BCTQ-FSS) on a 1–5 scale, pain visual analog scale on a 0–10 scale, conversion to open carpal tunnel release (CTR), complications, and reoperations. Outcomes were analyzed using a random-effects meta-analysis model. Metaregression was used to determine the association of patient- and study-level factors with ECTR outcomes.P < .001 for all). In metaregression, the strongest predictor of improvement in Q-DASH, BCTQ-SSS, and BCTQ-FSS was a greater preoperative score for that variable , indicating that patients with worse symptoms improved the most. The risks of conversion to open CTR, complications, and revision CTR were 0.7%, 0.7%, and 0.5%, respectively.A total of 17 studies with 1,632 patients treated with ECTR were included. Median follow-up durations ranged from 4 to 7 months depending on the outcome. Statistically significant and clinically important improvements were noted after ECTR for Q-DASH, BCTQ-SSS, BCTQ-FSS, and pain visual analog scale scores, with mean differences from baseline of −28.8, −1.8, −1.5, and −5.1, respectively (In a best-evidence synthesis of contemporary studies, ECTR resulted in significant improvements in function and pain, with a low risk of conversion to open surgery, complications, and reoperations over short-term follow-up.Patients treated with ECTR can expect generally favorable clinical outcomes over the short term. However, long-term outcomes after ECTR are not well characterized. The single-portal ECTR technique was first developed by Okutsu et al,Numerous meta-analyses of randomized controlled trials have compared the safety and effectiveness of ECTR and OCTR.www.researchregistry.com (reviewregistry1649). The methods and reporting adhered to the Preferred Reporting Items for Systematic Reviews and Meta-AnalysesThis contemporary best-evidence systematic review and meta-analysis was prospectively registered at https://www.jhsgo.org.The inclusion criteria for this best-evidence review required studies to be prospective, treat at least 50 hands with ECTR, report at least one outcome specified in this review within the 5-year postoperative follow-up period, and be published between January 2013 and January 2023, regardless of language. Studies were excluded if they reported outcomes from various CTR techniques, had ECTR performed with concomitant surgical procedures, had revision ECTR performed, or were published as abstracts or duplicate publications. The rationale for these best-evidence criteria is provided in A systematic search was conducted across multiple electronic databases, including MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials. The search strategy was purposely broad to ensure maximum search sensitivity. We used the terms [ECTR or endoscop∗] and [carpal or CTS or median nerve] for the MEDLINE searches. The search strategies were adapted as necessary for other databases. We also conducted supplementary manual searches in the Directory of Open Access Journals and Google Scholar and in the reference lists of included articles and relevant review articles. Two researchers with expertise in systematic reviews independently screened articles for eligibility. All potentially relevant studies were obtained in full text and reviewed. Discrepancies in study eligibility were resolved through discussion and consensus. The final search was conducted in February 2023.The researchers independently extracted data from eligible studies using standardized data collection forms. Data extraction discrepancies were resolved through discussion and consensus. We evaluated the methodological quality of studies using the National Institutes of Health assessment tool for before–after studies.I2 statistic where a value of 0% represented no heterogeneity, and larger values represented increasing heterogeneity.I2 > 50%). The independent variables of interest were age, sex, the preoperative value for patient-reported outcomes, the number of surgical incisions, follow-up duration, and sample size.Continuous outcomes were analyzed using the mean difference from baseline and 95% confidence interval (CI) for individual studies and the overall estimate. Binary outcomes were reported using the event rate and 95% CI. The overall results were obtained using a restricted maximum likelihood random-effects meta-analysis to account for heterogeneity among studies. In studies that reported outcomes from multiple ECTR subgroups, we combined the data; hence, each study provided only one ECTR group for each outcome.https://www.jhsgo.org). The median age of participants was 54 years (with a study-wide mean range of 43–65 years), with a higher proportion of female patients , and a median CTS symptom duration of 24 months , validating the utility of the best-evidence study selection criteria.Of the initial 536 titles and abstracts identified in the searches, 333 were deemed irrelevant and subsequently excluded. Among the remaining studies, the most common reasons for exclusion were insufficient sample size (48), review article 41), retrospective study design 38), studies that did not use ECTR or reported outcomes from various CTR techniques (19), and database analyses (19). Ultimately, the meta-analysis included 17 studies,, studies, retrospP < .001), and significant heterogeneity was identified among the studies , with significant heterogeneity identified among studies , and significant heterogeneity was observed , with significant heterogeneity identified among studies . Postsurgical complications were reported in 0.7% (95% CI: 0.2% to 1.2%) of the cases over a median follow-up duration of 7 months (https://www.jhsgo.org). The most common complication was pain/paresthesia (10 cases), all of which resolved within 6 months. Only one (0.1%) case of median nerve branch transection was reported,https://www.jhsgo.org). Heterogeneity among studies was negligible for all complication outcomes (I2 ≤ 5%).The short-term safety outcomes with ECTR were favorable. Conversion to OCTR was performed in 0.7% (95% CI: 0.3% to 1.2%) of the cases (P < .001), −1.9 to −1.7 for BCTQ-SSS , −1.5 to −1.3 for BCTQ-FSS , and −5.8 to −4.6 for pain VAS . As with the primary analysis, the overall mean difference and the entire 95% CI in the one-study-removed sensitivity analysis exceeded the MCID for each of these outcomes. Similarly, the overall event rates were comparable in the one-study-removed sensitivity analyses with values ranging from 0.6% and 0.9% for conversion to OCTR, 0.6% to 1.1% for complications, and 0.4% to 0.5% for reoperations. Three outcomes met the criteria for inclusion in the metaregression based on the number of studies and the presence of heterogeneity: Q-DASH, BCTQ-SSS, and BCTQ-FSS. For each of these outcomes, the baseline value was the strongest predictor of postoperative change, where more severe symptoms were associated with a greater magnitude of postoperative improvement (https://www.jhsgo.org).The trim-and-fill analysis resulted in identical estimates as the main analysis, except for reoperations that were 0.5% in the main analysis and 0.4% in the trim-and-fill analysis. Thus, publication bias was negligible for all outcomes, suggesting that the meta-analysis results were not influenced by the selective publication of studies based on the direction or strength of their findings. Additionally, no individual study substantially influenced the overall conclusions of the meta-analysis. By rerunning the meta-analysis excluding each study at a time, the overall mean differences ranged from −30.7 to −24.3 for Q-DASH . Fourth, the outcomes reported in this meta-analysis were observed over relatively short follow-up durations, ranging from 4 to 7 months. Finally, most of the recent ECTR literature consists of studies of lower methodological quality as evidenced by the large number of studies excluded from this review due to retrospective designs and small sample sizes. Overall, although the short-term outcomes with ECTR appear favorable based on the best evidence in the literature, the overall quality of available ECTR research is suboptimal, underscoring the need for larger prospective studies to substantiate these findings over longer follow-up durations.The findings of this systematic review and meta-analysis are consistent with those of the American Association of Orthopedic Surgeons that stated “limited evidence supports that if surgery is chosen, a practitioner might consider using endoscopic carpal tunnel release based on possible short-term benefits.”,,Despite the methodological limitations of the literature, ECTR utilization has been increasing, particularly in the Medicare population.,,,,Several limitations are associated with this review. First, although the results presented here were developed using best-evidence synthesis methods, they represent level IV evidence. Only two randomized trials compared ECTR with another CTR technique that met the best-evidence criteria,In a best-evidence synthesis of contemporary studies, ECTR resulted in significant improvements in functional outcomes and pain relief, with a low risk of conversion to open surgery, complications, and reoperations over short-term follow-up.


Glioblastoma (GBM) is among the most fatal and recurring malignant solid tumors. It arises from the GBM stem cell population. Conventional neurosurgical resection, temozolomide (TMZ)-dependent chemotherapy and radiotherapy have rendered the prognosis of patients unsatisfactory. Radiotherapy and chemotherapy can frequently induce non-specific damage to healthy brain and other tissues, which can be extremely hazardous. There is therefore a pressing need for a more effective treatment strategy for GBM to complement or replace existing treatment options. Cell-based and cell-free immunotherapies are currently being investigated to develop new treatment modalities against cancer. These treatments have the potential to be both selective and successful in minimizing off-target collateral harm in the normal brain. In this review, several aspects of cell-based and cell-free immunotherapies related to GBM will be discussed. According to the 2016 World Health Organization (WHO) histopathological and clinical criteria, gliomas are classified as grades I-IV . In contGlioblastoma (GBM) has been described as a grade 4 tumor by the WHO and is among the most fatal and recurring malignant solid tumors to date  accountiAs early as the mid-nineteenth century, it was proposed that cancer treatment could be achieved by modulating the body’s immune system to combat cancer . Its dis22.1in vivo persistence (Chimeric antigen receptor (CAR) T cell therapy holds one of the most promise as an anti-cancer therapeutic technology. CAR are synthetic molecules formed by four regions, the antigen recognition structural domain , the extracellular hinge region, the transmembrane structural domain (consisting of a hydrophobic α-helix across the cell membrane) and the intracellular T cell signaling structural domain   intendedsistence . Third gsistence . Fourth sistence . The fifsistence Figure 2CAR-T cells were first used to treat hematological malignancies and have shown remarkable efficacy , 22. For2.1.1IL-13Rα2 as a target for CAR-T in the treatment of GBM. It’s a membrane receptor with a high affinity for the anti-inflammatory cytokine interleukin 13 and it has been discovered to be overexpressed in a variety of solid tumors, most notably GBM, and has been related to poor prognosis . IL-13Rαin situ mouse models, this approach has not yet been validated in patients. In 2015, they published the first promising human clinical study of intracranially administered IL13Rα2-specific CAR-T cells for GBM, which set the stage for the future application of improved peripatetic CAR-T cells therapy , as the first tyrosine kinase receptor to be cloned, is still at the leading edge of targeted cancer therapy. Being the most common variant of EGFR, EGFRvIII is usually expressed in GBM  and is ain situ models of human EGFRvIII+GBM, EGFRvIII-targeted CAR T cells were also able to suppress tumor development. They also planned a phase I clinical research utilizing humanized scFv-transduced CAR T cells targeting EGFRvIII in patients with residual or recurrent GBM based on these findings   targeting EGFRvIII mutant oncoproteins is a therapeutic approach , and sec2209376) . A first2209376)  in whichlization , hence tlization  used a t2.1.3in vitro with the aim of identifying and killing EphA2-positive glioma cells and glioma-initiating cells, as well as inducing tumor induction in an in situ xenografted GBM with Severe Combined Immunodeficiency (SCID) mouse model of regression. However, such manipulations are still carried out through highly artificial means and may not better predict future clinical effectiveness. H.T. Lin et al.  for broader application. Trivalent CAR-T cells have the potential to overcome antigen heterogeneity in GBM and improve treatment results. Furthermore, Joseph H. Choe et al.  as a delivery vehicle based on NEs to effectively deliver PTX to tumour cells and induce cytotoxicity and inhibit post-operative recurrence of GBM , the most prevalent leukocyte population in the blood, may be rapidly recruited to sites of inflammation, and are thought to be a powerful platform for tumour-targeted drug delivery, similar to mesenchymal stromal cells (MSCs) . More ime of GBM . Furthere of GBM  internale of GBM –63. The henotype . To addrhenotype  mounted 2.3MSCs are pluripotent stem cells, which are normally derived from bone marrow, umbilical cords/placenta, and adipose tissue. MSCs have tissue healing capability and low immunogenicity, and they are not restricted by the BBB/BTB, so they can be intrinsically subsumed into the brain tumour site , 65, ove2.3.1in vivo situation where tumour cells are integrated with supportive cells more unknown. Improving the efficiency of MSCs in the treatment of GBM requires the use of appropriate tools and technical abilities. Extensive research has revealed the promise of suicide genes in the treatment of glioma tumors. Enhancing their effectiveness relies on the ability to apply the right tools and techniques. Saeed Oraee-Yazdani et al.  in GBM treatment affected by the rapid clearance and non-specific bio-distribution of gadolinium-based drugs, Yen-Ho Lai et al.  and chemokines  to better recruit MSCs to GBM sites , 98. In on model . OV seleon model . The useon model . Delta-2on model . MSCs loon model . MSCs enon model .2.4in vitro destruction by healthy allogeneic NK cells. Their findings demonstrated that GBM tumour-infiltrating NK cells acquired an altered phenotype associated with impaired lytic function when compared to matching peripheral blood NK cells from GBM patients or healthy donors. This immune evasion approach was attributed to αv integrin-mediated TGF-β activation, which directed interactions between GSCs and NK cells. In contrast, blocking the αv integrin/TGF- β axis can increase NK cell antitumor function. Gregory J Baker et al.  cells play an essential role in the body’s anti-infection, anti-tumour, and immunomodulatory processes as recognition and effector cells in the innate immune system. NK cells do not inhibit the killing of their own normal cells, but selectively recognize and kill cells that are low in expression or lack their own major histocompatibility complex (MHC)-I molecules . MHC-I mr et al.  found thin vitro and in vivo. In contrast to CAR-T cells, which require autologous cells for each patient, NK cells are safe under allogeneic circumstances, which broadens the pool of cell donors capable of producing therapeutically meaningful amounts of CAR-NK cells for therapy  are now recognized as the most potent and only specialist antigen-presenting cells in the body capable of activating naïve T cells. It is called after the numerous dendritic protrusions that emerge from the cell surface during maturation. Immature and mature dendritic cells perform distinct tasks, with immature dendritic cells being good at antigen differentiation and phagocytosis and mature dendritic cells being good at antigen presentation. DC have MHC class I and II molecules on their surface, which when combined with antigenic peptides trigger CD4+ helper T cells (Th) and CD8+ cytotoxic T lymphocytes (CTL) to elicit specific anti-tumour cellular and humoral immunity, inhibiting tumour cell growth . DC alsoClinicalTrials.gov, NCT01280552). HLA-A2 primary tumour antigen expression was more frequent than in HLA-A1 patients. HLA-A2 patients had a higher immune response (by Elispot) and patients in the pre-specified subgroups of methylated and unmethylated MGMT achieved meaningful therapeutic benefit with ICT-107. This was the first vaccination study to demonstrate a clinical benefit in GBM, and it paved the way for a phase III trial in patients with HLA-A2+ newly diagnosed GBM. Linda M Liau et al.   Table 2.0045968) . The use0045968) , 134. Th2.6via the impaired BBB/BTB, and both cells are converted into critical drivers of tumour development by acting as TAMs together to infiltrate at GBM locations (Microglia and tumour-associated macrophages (TAMs) are the main components of GBM myeloid cells, which are maintained by self-renewal under physiological conditions and are associated with functions such as CNS inflammation and development . Under pocations .TAMs are the most common immune cells in the TME, and their phenotype is diverse and flexible . The bul33.1Despite the many advantages of MSCs in the fight against GBM, there are some limitations to the use of MSCs, such as low biological activity and limited accessibility , 80, 87.MSCs can be an abundant source of Exo, and all Exo express the same group of proteins, such as tetraspanins , adhesion proteins , Alix and TSG101 –150. ExoMSCs exosomes (MSCs-Exo), as paracrine mediators of the therapeutic effect of MSCs, have comparable biological activity to MSCs. However, compared with MSCs, MSCs-Exo are smaller, penetrate biological membranes more easily, are less immunogenic, more biocompatible, and better preserved , 156. Pr3.2via several mechanisms. At first, viruses proliferate in tumour cells and directly lyse tumour cells  have become effective new therapeutic tools in this field –182. OV ur cells . Then, lur cells –191. Dueur cells Table 3.3.2.1Oncolytic herpes simplex virus (oHSV-1) is a neurophilic double-stranded DNA virus. Typically, wild HSV-1 is neurotoxic, so the virus must be genetically modified or greatly attenuated to ensure safety. After genetic modification, OV can still maintain its ability to reproduce while replicating specifically in tumour cells, and therefore OV is widely exploited. Based on previous findings, oHSV was the first state-of-the-art genetically engineered OV to be licensed by the United States FDA for cancer treatment  and was in vivo.However, as GBM is a primary brain tumour of the human central nervous system, more attention deserves to be paid to the safety of oHSV-1 in the fight against GBM. In past studies, oHSV-1 is encoded by the γ34.5 gene, an ICP34.5 protein, which is neurotoxic . To limiG207 is a second generation oHSV-1 that inactivates the ICP6 gene by deleting the double copy γ34.5 gene while inserting the lacZ gene at the UL39 locus. During a Phase I clinical trial of genetically engineered oHSV-1 G207 by GK Friedman et al. , oHSV-1 G47Δ is a third generation oHSV-1 with a triple mutation in α47 deleted from the G207 genome. Compared to G207, G47Δ is further attenuated in normal cells, but has enhanced efficacy in anti-tumour, as well as a greater safety profile. oHSV-1 G47Δ showed efficacy and safety in GBM was confirmed by the American Association for Cancer Research in 2016 . In subs3.2.2Adenovirus is a non-enveloped virus with an icosahedral capsid containing approximately 38 kb of genomic double-stranded DNA. There are more than 50 serotypes of adenovirus in humans, of which types 2 and 5 have been most frequently studied for the manufacture of lysing viruses , 206. Invia the integrins avβ3 and avβ5 into tumour cells, which further enhances tumour targeting  that degrades HA and spreads efficiently in the tumour. It has the same mechanism of action as Delta-24-RGD, ICOVIR17 deletion of 24 base pairs in the Rb-binding domain of E1A for tumour-selective replication and RGD modification in the fibril to amplify tropism, except for two additional modifications: insertion of an E2F binding site in the E1A promoter and insertion of the SPAM1 gene encoding PH20 HA after the fibril, which is controlled by the major late promoter control . Normallin vivo. Jawad Fares et al.  , 217. Ths et al.  conducte3.2.3Reovirus (RV) belongs to the wild-type OV family of eutheroviruses and is characterized as a staged double-stranded RNA virus. The three prototypical serotypes of RV, first identified in the 1960s, are well characterized as serotype 1 Lang (T1L), serotype 2 Jones (T2J) and serotype 3 Dearing (T3D) . RV is w3.3GBM generates an immunosuppressive environment through multiple mechanisms, including the programmed cell death protein 1 (PD-1), cytotoxic T lymphocyte antigen 4 (CTLA-4), lymphocyte activating gene 3 (LAG-3) pathway . AlthougThe immunosuppressive properties of the GBM microenvironment lead to immune evasion by tumour cells, making inhibition of immune checkpoints such as PD-1 alone ineffective , 227. PD3.3.16-methylguanine-DNA methyltransferase (MGMT) renders tumors resistant to TMZ, MGMT promoter status predicts both prognosis and therapeutic response to TMZ chemotherapy. Antonio Omuro et al.  of HSV-1 against PD-1, which also expresses a single-chain fragment mutable antibody (scFvPD-1). Irene Appolloni et al. , a triple combination that could assist in curing less immunogenic malignancies such as GBM (31). Also based on the fact that OV and PD-1 inhibitors have become standard immunotherapies against certain cancers, Carmela Passaro et al.  conductei et al.  also inv4There is a deepening perception of what cell-based and cell-free immunotherapy can bring to GBM, which offers new hope for GBM patients, but many details and questions remain to be explored and elucidated. According to recent statistics, adjuvant immunotherapy can prolong survival and significantly improve outcomes for patients with recurrent GBM compared to those treated with only surgery, radiotherapy or chemotherapy. To summarize the lessons learned, the extremely specific and heterogeneous nature of GBM, as well as the complicated immune resistance mechanisms and immunosuppressive TME have resulted in relatively limited response effectiveness and durability of response to treatment drugs. The BBB/BTB is another barrier to medication delivery. These are still tough breakthroughs in GBM therapy. We not only obtained some novel findings on the theoretical study of the local immune characteristics of glioma, but we also provided an experimental basis for the comprehensive diagnosis and treatment of regulating and intervening in the immune microenvironment of glioma. More significantly, “supporting the righteousness” and improving the immune microenvironment, in conjunction with “elimination of evil” by anti-tumour cells, will ideally decrease disease mortality rates, extend patient life, and genuinely help patients. In fact, the treatment of GBM is actually a complex “project” and satisfactory results can hardly be achieved with only one treatment. Immunotherapy research for GBM should be coupled with other treatment modalities in future anti-GBM research, resulting in truly tailored and complete care strategies for patients. It is recommended to develop combined treatment techniques based on immunotherapy, molecular targeted therapy, and radiation to maximize therapeutic efficiency and reduce acquired immunotherapy resistance. The industry therefore needs to innovate, integrate and translate to drive immune combinations forward in a sustained manner. Secondly, assessing therapeutic response to immunotherapy is difficult, and in the future, a standardized imaging and molecular biology evaluation system will be required to reflect and forecast patient outcomes. Cell-based and cell-free immunotherapy are predicted to become an essential component of future glioma treatment when these challenges are solved.MW conceived this article. MW and XW drafted the manuscript. MW and XW drew the illustrations and tables. MW, XW, XJ, JZho and YZ revised the article. MW, YL and JZha checked and edited the article. YL and YY supervised the article. All authors have read and agreed to the final version of the manuscript.


Hashimoto's thyroiditis (HT) is an autoimmune disorder affecting the thyroid gland and may present as goiter or atrophic thyroiditis that may result in various metabolic and inflammatory disorders. The aim of this study is to determine the changes in serum levels of interleukin-17 (IL-17), IL-23, neopterin, and nesfatin-1 parameters in HT patients and to evaluate the possible relationship among these parameters.90 HT patients and 30 healthy individuals were included in this study. Demographic data of the patients included in the study were recorded and detailed physical examinations were performed. IL-17, IL-23, neopterin, and nesfatin-1 levels were measured in the serum samples of the participants by the ELISA method. Hashimoto’s thyroiditis (HT) is one of the most common and organ-specific autoimmune diseases and it’s caused by the loss of immune tolerance of the thyroid gland Identification of a new subtype of T helper cells producing the IL-17 modified model of the Th1-Th2 paradigm and termed Th17 cells. The latter have high ability to stimulate acute and chronic inflammation making them an important role in the development of autoimmune disorders IL-23 is an inflammatory cytokine and has been found to be essential for disease development in various autoimmune disease models Neopterin is produced by macrophages and responsible for activation the cellular immune response Nesfatin-1 is a neuropeptide produced in the hypothalamus and suppresses hunger feeding and increases insulin secretion from pancreatic beta islet cells. Therefore, nesfatin-1 has attracted attention as a new therapeutic agent in the treatment of obesity and diabetes mellitus, especially in thyroid diseases Since IL-17 and IL-23 are inflammatory cytokines, high concentrations of these markers arethought to be associated with the severity of HT and the progression of the disease, based on the autoimmune association between thyroid hormones, TPO, nesfatin-1 and neopterin. This study reveals whether these biomarkers can be used in patients with HT. Since HT remains a common disease with unknown pathogenesis, we aimed to determine the levels of these biomarkers in serum samples of patients with HT.A total of 120 people, 90 patients who were diagnosed with HT by the Department of InternalMedicine, Harran University Faculty of Medicine, and 30 healthy people were participated in our study. Demographic data of the patients included in the study were recorded and detailed physical examinations were performed. The study was approved by the scientific ethics research coordinatorship of Harran University with the protocol number HRU/21.06.35 dated 15.03.2021.The patients included in our study were divided into three groups. The first group consisted of 30newly diagnosed patients (20 men and 10 women) who had never been treated for HT. The mean age of these patients was 36.75±11.93 years. The second group consisted of 30 patients  diagnosed with euthyroid HT and not treated with thyroid hormones before. The mean age of the patients was determined as 33.14±9.35 years. In the third group, there were 30 HT patients (9 men and 21 women) who were treated with levothyroxine, their mean age was 35.48±11.64 years. In addition, a healthy control group was formed with 30 individuals (12 men and 18 women) who did not have any autoimmune and chronic diseases. The mean age of these individuals was 39.00±8.46 years.Thyroid stimulating hormone (TSH), free T3 , free T4 , thyroid peroxidase antibody (anti-Tpo), thyroglobulin antibody (anti-Tg), neopterin, nesfatin-1, IL-17 and IL-23 levels were measured in the serum samples of the participants.Venous blood was drawn for laboratory tests after 8-12 hours of fasting for each participant. Measurements of the parameters from serum samples were determined by using Atellica  instrument. Analyses were performed according to the manufacturers’ instructions for each ELISA kit. Optical density was read at 450 nm. The cutoff value of each assay was calculated according to the manufacturer’s instructions.Statistical analyses were carried out using the SPSS software 26.0 and Minitab 20.3. . Descriptive statistics were provided as min, max, and meanstandard deviation. The Shapiro-Wilk test was preferred to check normality. Levene’s test was used to determine if the samples have equal variances. The Welch’s test for one-way ANOVA combined with the Games-Howell multiple comparison post-hoc test to perform pairwise comparisons across different groups was used. Cor relations among variables were determined using Pearson correlation. The values which are greater than 0.05 were considered not statistically significant.The serum TSH, FT3, FT4, anti TPO, anti TG, neopterin, nesfatin-1, IL-17 and IL-23 levels of 120 individuals participating in the study are shown in Compared to the control group (30.73±4.63), IL-17 levels were found to be higher (p<0.001) in the hypothyroidism HT, euthyroid HT, and HT patients treated with levothyroxine groups . Likewise, IL-23 levels were found to be higher in hypothyroidism HT (44.46±8.40 pg/mL) than the control group, euthyroid HT and HT patients treated with levothyroxine groups  (p<0.001) .Similaly, serum neopterin levels were found to be significantly high in hypothyroidism HT (p 0.001) but its levels in controls, euthyroid HT and HT patients treated with levothyroxine groups were as follows:  Significantly lowest level of nesfatin-1 was found in HT whereas its levels in controls, euthyroid HT and HT patients treated with levothyroxine and hypothyroidism HT, groups were  p<0.001, .Patients with high TSH levels were divided into three groups according to their symptoms and free T3 and free T4 levels. Newly diagnosed HT with hypothyroidism; TSH levels were above normal >10 mIU/L, and TPO and TG antibodies were positive.Patients with subclinical hypothyroidism (euthyroid) HT and thyroid replacement therapy with levothyroxine, have normal TSH levels (<10 mIU/L), and TPO and TG antibodies are negative in these two groups of patients .The Welch’s ANOVA test revealed statistically significant differences between the four groups’means for neopterin, nesfatin, IL-17, and IL-23.  = 650.9, p < 0.001; nesfatin- F  = 195.55, p < 0.001, IL-17 - F  = 369.96, p < 0.001, IL-23- F  = 353.26, p < 0.001), indicating that the average hormone values measured in groups were not equal.The Games-Howell Post-Hoc analysis results were presented in Our study showed that both serum levels of IL-23 and IL-17 were significantly increased in HT patients with newly diagnosed HT hypothyroid patients, euthyroid HT patients, and HT treated with levothyroxine patients respectively compared to the healthy control group.Variables including neopterin, nesfatin-1, IL-17, IL-23, TSH, FT3, FT4, Anti-TPO, and Anti-TG were recruited into correlation analysis based on each group. Correlations of all variables were determined using Pearson correlation. No statistically significant correlation was found between any of these variables in the hypothyroidism group. The correlation coefficient between interleukin-17 and TSH was about r=-0.426 (p=0.021) indicating that there was a moderatenegative correlation in the euthyroid HT group. The correlation coefficient between neopterin andanti-TPO was about r=0.413 (p=0.026) indicating that there was a moderate positive correlation in the HT treated with levothyroxine group. In the same group, correlation coefficient between TSH and anti-TPO was also found significant r=0.519 (p=0.004). In the healthy control group, three statistically significant correlations were found. First correlation was between neopterin and nesfatin-1, r=0.427 (p=0.019) indicating that there was a moderate positive, as neopterin increases nesfatin-1 increases. Second correlation was between interleukin-23 and anti-TG, r=0.391 (p=0.033). Third correlation was between TSH and anti-TG, r=0.484 (p=0.007), The relationship between neopterin and nesfatin-1 in the healthy control group shown in HT is a chronic autoimmune disease with a complex and heterogeneous course that primarily causes destruction and dysfunction of the thyroid gland In HT, accumulation of lymphocytes in the thyroid gland eventually leads to thyroid fibrosis and gradual tissue destruction Recently, it has been suggested that Th3 and Th17 cells play an important role in the pathogenesisof chronic inflammatory diseases, including HT, chronic obstructive pulmonary disease (COPD), diabetes, and rheumatoid disease In our study, the increase in serum IL-23 levels in all three HT patient groups reflects that this IL-23has a role not only in the initiation of pathogenic processes but also in the maintenance of autoimmune inflammation. The correlation established between IL-17 and IL-23 for euthyroid patients confirms the positive association between these two cytokines and their involvement in disease onset. Our results showed significant differences in serum levels of IL-17 between healthy control and all HT patients. It decreased in patients with hypothyroid HT, which increased during treatment with levothyroxine, suggesting that IL-17 levels may be affected by hypothyroidism. Decreased IL-17 levels in hypothyroidism may be associated with depression of humoral and cell-mediated immunity in this functional state. These changes in serum levels of IL-17 were developed on the basis of general immune disorders found in the Th17 pathway in HT It is well known that neopterin is proinflammatory marker synthesized by macrophages upon stimulation with gamma interferon-gamma and is indicative of cellular immune system activation. Levels of neopterin are elevated in conditions which has an immunological component such as autoimmune disease, viral and bacterial infections and malignancy Nesfatin-1 is satiety molecule produced in hypothalamus. It participates in the regulation of appetite and hunger Our results suggest that the role of IL-23, IL-17 axis may play a role in the etiopathogenesis and development of HT in all stages of the disease as cause or effect. Moreover, our data show that the importance of IL-23 is more pronounced than IL 17 in the development and severity of HT. Increased neopterin production is also associated with HT and its levels correlate with the extent and the activity of the disease. Nesfatin-1 causes satiety. People with underactive thyroid gland would gain weight, and we always attribute this to slowing down of metabolism. Weight gain in this period may be related to increased appetite and may be due to low nesfatin-1 levels in these patients. This shows us that patients with Hashimoto’s thyroiditis are prone to obesity even if thyroid hormones do not decrease. Second, even if we correct the patient’s hormones, nesfatin-1 remains low compared to the health for these purposes. Further studies with better design are needed to be done on this subject.All authors contributed to the writing of the initial draft. NB contributed to the discussion and edited the manuscript. NB and AÖ reviewed the manuscript. NB did the practical work in the lab. All authors have read and agreed to the published version of the article.Written consent has been obtained from each patient or subject after full explanation of the purpose and nature of all procedures used.This research was supported by the scientific research coordinatorship of Harran University, Turkey (Project no 21179).The study was approved by the scientific ethics research coordinatorship of Harran University with the protocol number HRU/21.06.35 dated 15.03.2021.All the authors declare that they have no conflict of interest in this work.


The relationship between cyber video game addiction and impulsivity is controversial. Some studies have shown a significant link, others have found no association between the two behaviours.To study impulsivity in adolescents with problematic use of internet video games.This was a cross-sectional, descriptive and analytical study, conducted among a sample of adolescents randomly collected in 6 schools in the region of Sfax-Tunisia, during the month of February 2022. The rate of addiction to video games was assessed by the 20-item “Internet Gaming Disorder-20” (IGD-20) scale and impulsivity by the 30-item “Barratt Impulsivity Scale” (BIS-11). Both scales are validated in Arabic.The study involved 360 secondary school students, with a mean age of 16.62 +/- 0.822 years. The sex ratio was 1.09.A gaming addiction was found in 4.7% of cases.Similarly, impulse control disorder was noted in 23.6% of adolescents.Problematic internet game use was significantly related to motor impulsivity (p=0.025).There was no significant association between cyber video game addiction and cognitive or non-planning impulsivity.According to the results, impulsiveness is a factor to consider for understanding the development of addiction to internet video games. Thus, impulsiveness should be taken into account to explain problematic gaming behaviour as well as to design preventive and treatment interventions.None Declared


Increased triglycerides (TGs) are a major risk factor for cardiovascular disease. Furthermore, hypertriglyceridemia is commonly associated with a reduction of high-density lipoprotein cholesterol (HDL-C) and an increase in atherogenic small-dense low-density lipoprotein (LDL-C) levels. Studies provide support that polyunsaturated omega-3 fatty acids (ω3-LCPUFAs) are cardioprotective and have antithrombotic and anti-inflammatory effects. The potential effects of ω3-LCPUFAs on cardiometabolic factors and anti-inflammatory actions in children with acute lymphoblastic leukemia (ALL) are limited. This is a secondary analysis of a previous clinical trial registered at clinical trials.gov (# NCT01051154) that was conducted to analyze the effect of ω3-LCPUFAs in pediatric patients with ALL who were receiving treatment.Objective: To examine the effect of supplementation with ω3-LCPUFAs on cardiometabolic factors in children with ALL undergoing treatment.Thirty-four children  aged 6.7 ± 2.7 years who were newly diagnosed with ALL were evaluated. Children were randomized to receive either ω3-LCPUFAs or placebo capsules (sunflower oil). ω3-LCPUFAs were administered in the form of 500-mg soft capsules. The ω3-LCPUFA capsules contained 225 mg of DHA, 45 mg of EPA, and 20 mg of another ω3-LCPUFAs. The omega-3 dose was administered at a rate of 0.100 g/kg of body weight/day for three months. Main outcomes: Fasting cholesterol, HDL-C, very-low-density lipoprotein (VLDL-C), TGs, atherogenic index of plasma (AIP), android/gynoid ratio (A/GR), IL-6, TNF-α, and percentage of fat mass (DXA) were measured in all patients. Fatty acid analyses in red blood cells were performed with gas chromatography.We found significantly lower levels of TGs (p=0.043), VLDL-C (p=0.039), IL-6 (p=0.025), and AIP (p=0.042) in the ω3-LCPUFAs group than in the placebo group at three months. In contrast, the total cholesterol concentration was higher at 3 months in the ω3-LCPUFAs group than in the placebo group . The number of children with hypertriglyceridemia  tended to be lower between the time of diagnosis and after 3 months of supplementation with ω3-LCPUFAs.These findings support the use of ω3-LCPUFAs to reduce some adverse cardiometabolic and inflammatory risk factors in children with ALL.ClinicalTrials.gov, identifier NCT01051154. AlthougDifferent studies have shown that patients with ALL presents a proinflammatory state at the time of diagnosis and during and after chemotherapy for up to 5 years after the start of treatment, characterized by increased levels of cytokines  . In thisOn the other hand, several studies suggest that dietary supplementation with ω3-LCPUFAs, such as DHA and EPA, can be beneficial for cancer treatment, as it decreases metabolic risk; modulates several aspects of the inflammatory response; decreases inflammatory markers, plasma levels of TGs and LDL-C; and increases HDL-C . NeverthHowever, to our knowledge, there are no randomized controlled clinical trials comparing the direct changes in the lipid profile and inflammation markers induced by ω3-LCPUFA supplementation and assessing adherence to treatment by gas chromatography (gold standard) in ALL patients.Therefore, we present results from a secondary analysis of data obtained in a randomized clinical trial that was previously conducted to evaluate the effect of supplementation with ω3-LCPUFAs on body composition in children with leukemia (under review).The aim of the present analysis was to examine the effect of supplementation with ω3-LCPUFAs on several cardiometabolic factors in children undergoing treatment for ALL with the hypothesis that ω3-LCPUFAs reduce inflammatory cytokine and TGs levels and increase HDL-C in these patients.22.1This analysis is part of a randomized clinical trial designed to evaluate the effect of supplementation with ω3-LCPUFAs on body composition in children with leukemia at the end of the remission phase and three months after supplementation, registered in the ClinicalTrials.gov database . This study was conducted in accordance with the Declaration of Helsinki, and it was approved by the National Committee of Scientific Research of Instituto Mexicano del Seguro Social (IMSS) and was conducted in the Unit of Research in the Medical Nutrition in a Pediatric Hospital in Mexico City . All parents or legal guardians of the children provided written informed consent prior to study entry.2.2We included 72 children with ALL at the Pediatric Hospital of the National Medical Center XXI Century IMSS, National Medical Center, and at the “Gaudencio González Garza” General Hospital, of the “La Raza” National Medical Center. Eligibility criteria were children newly diagnosed with ALL  at the start of their chemotherapy treatment. Children who had previously been treated with chemotherapy in another institution, those with severe comorbidities, using corticosteroids, taking fish oil supplements during the previous weeks, who were unable to swallow ω3-LCPUFA or placebo capsules, and those who reported allergies to fish intake were excluded. From 72 children with ALL, in this secondary analysis, only 34 children were included , due to participants dropping out for different reasons, such as discontinued intervention, death, clinical complications (neutropenic colon), or lack of adherence. We have reported this information in the manuscript of a clinical trial on body composition and omega 3 in children with ALL .2.3http://www.msaghaei.com/Softwares/dnld/RA.zip)  or to the ω3-LCPUFA group (intervention) by a computer-generated list of random numbers using software for parallel groups   The rand2.4®Kabi, “Frebini Plus” that covered 15% of their total energy expenditure, which was calculated by the Schofield formula. Each 236 ml container of the supplement contained 351 kcal (1491 kJ), 8.9 g of protein, 43.7 g of carbohydrates, and 15.7 g of lipid, and three flavors were employed . In those patients who presented secondary complications, such as diabetes and pancreatitis, supplementation was contraindicated, and therefore, they were excluded from the study. Children and their parents were instructed to register capsules and oral supplements at the beginning of chemotherapy. During the study period, supplementation was supervised by phone or in the hospital if the patients were staying there by medical personnel.Children received either capsules of ω3-LCPUFAs or placebo capsules. ω3-LCPUFAs were administered in the form of 500 mg soft capsules of natural TGs, made from gelatin, formulated without artificial colors or flavors, molecularly distilled, and of pharmaceutical grade . The omega dose was administered at a rate of 0.100 g/kg of baseline body weight/day. The ω3-LCPUFAs comply with the principles established for fats according to the European Pharmacopoeia Standard (EPS) and according to the Council for Responsible Nutrition (CRN) and the Global Organization (CRNGO). Consequently, ω3-LCPUFAs are a safe product that does not exceed the maximal allowances for contaminants such as peroxides, heavy metals, dioxins, and PCBs. The placebo capsule contained 500 mg of sunflower oil . All capsules contained vitamin E to act as an antioxidant. The odor and appearance of the ω3-LCPUFA capsules and the placebo capsules were comparable, and both were strawberry-flavored to mask their taste. During the study time, all the participants were provided with an oral supplement of the brand Fressenius2.5When the patient was discharged, compliance was monitored by the leftover pill count at their next appointment. In addition, the concentration of polyunsaturated fatty acids in erythrocyte membranes was determined before and during intervention with ω3-LCPUFAs. All side effects that the children presented during the intervention with ω3-LCPUFAs, or placebo were documented and registered by one of the researchers.2.62.6.1th to 94th percentile, and underweight as < 5th percentile)  with the subjects wearing lightweight clothing. Height was measured to the nearest 0.1 cm with a wall-mounted stadiometer . BMI percentiles for age and sex were calculated according to the Centers for Disease Control (CDC) normative curves using the computer software Epi-info  . All mea2.7th percentile, and >95th percentile have been recently associated with higher metabolic risk in children  with commercially available kits. TGs levels were considered acceptable <75/<90 mg/dL, borderline 75-99/90-129 mg/dL, and high ≥100/≥130 mg/dL for children aged < 10 and ≥ 10 respectively. HDL-C levels were considered low at <40 mg/dL ; total cchildren .Inflammatory markers such as IL-6 and TNF-α were determined in duplicate using high-sensitivity enzyme-linked immunosorbent assay (ELISA) kits, according to the manufacturer’s instructions . All assays were carried out in duplicate; coefficients of variation were 8% for ELISAs.2.8Analyses were performed with a 7820A gas chromatograph  with a flame ionization detector (FID) as described previously 2.9p < 0.05 were considered statistically significant.The data were analyzed using SPSS 21.0 software for Windows . Data are presented as the mean ± standard deviation (SD) or as the median , according to data (determined by the Shapiro-Wilk test), while categorical variables are presented as frequency (percentages) and were analyzed by Pearson’s chi-square test and Fisher’s exact test as appropriate. The crude significance of within-group and intergroup differences was tested by Student’s t test, paired-samples t test, the Wilcoxon test, or the Mann–Whitney U test, as appropriate. Values of 3We summarize the Consolidated Standards of Reporting Trials followed in this study in 3.1p = 0.039) and TGs (p = 0.043) in the ω3-LCPUFA group than in the placebo group at three months. In this sense, after follow-up, we showed that the ω3-LCPUFA group had significantly decreased levels of TGs and VLDL-C between diagnosis and at 3 months of intervention compared with the placebo group  (p = 0.009) in the ω3-LCPUFA group than in the placebo group. We found that 80% (16/20) and 85% (12/14) of children had hypertriglyceridemia in the placebo group and the ω3-LCPUFA group, respectively, at the time of ALL diagnosis. In addition, when analyzing each group, the VLDL-C concentrations decreased significantly between the time of diagnosis and after 3 months of supplementation with ω3-LCPUFAs (p = 0.009), and the number of children with hypertriglyceridemia (p = 0.054) tended to be lower. The AIP at 3 months was significantly lower in the ω3-LCPUFA group than in the placebo group ; The number of children with an A/GR > 95th percentile was higher in the placebo group than in the ω3-LCPUFA group (47% p =0.028).Blood lipid profiles for each group at baseline and after 3 months of intervention are shown in ctively) 3. In cop = 0.025).Inflammatory markers in children with ALL from baseline and after three months of supplementation with placebo or ω3-LCPUFA capsules are shown in p = 0.033), which was consistent at 3 months of supplementation. At three months, the alpha-linolenic acid (ALA) concentration was lower in the ω3-LCPUFA group than in the placebo group . After the administration of ω3-LCPUFAs, there were significant and progressive increases in the concentrations of DHA  and EPA (0.28 vs. 1.11 w/w% (p <0.0001) between baseline and at 3 months. The concentrations of DHA and EPA in the ω3-LCPUFA group increased by 193% and 289%, respectively.th percentile was significantly lower in the ω3-LCPUFA group than in the placebo group during the administration of ω3-LCPUFAs. These results confirm that the intervention was effective because of the variation in the rate of change in erythrocyte membranes after supplementation with ω3-LCPUFAs.In this secondary analysis, we confirmed the effect of ω3-LCPUFA supplementation at a rate of 0.100 g/kg/day on the lipid profile, specifically on the TGs and VLDL-C concentrations during the first 3 months of ALL treatment. We observed a significantly lower concentration of TGs and VLDL-C in the ω3-LCPUFA group than in the placebo group. We found that 82.4% (28/34) of the children had hypertriglyceridemia and low levels of HDL at the time of ALL diagnoses. These results are consistent with previous findings from ALL studies –32. In ap = 0.025) and 113 (p = 0.016) and in the total cholesterol concentration at day 113 (p = 0.027) in the intervention group compared to 22 historical controls. Nevertheless, when the authors corrected for one extra PEG-asparaginase dose, the level of significance in both lipid levels was reduced. In this study, the authors did not discuss the bias that the results could present in the intervention group because three patients presented a percentage of compliance of less than 80% . However, they recognized their limited sample size and the disadvantage of using historical controls, and the fact that it was not a randomized parallel-group trial. In our study, both groups were randomized, and the sample size, although small, was higher than that in Lauman’s study.There is considerable evidence that EPA and DHA have independent effects on multiple cardiometabolic risk factors, including blood pressure and cardiac function and lipids, as well as anti-platelet, anti-inflammatory, pro-resolving, and antioxidative actions . To our On the other hand, in a retrospective study, Salvador et al.  evaluatep = 0.009). These results contrast with those reported by Laumann and Salvador . In a longitudinal study (p = 0.002) and in the percentage of patients with a high index (> 0.24) after finishing 3 months of follow-up.The AIP is a biomarker of atherogenic dyslipidemia that, through non-HDL-C and/or the TG/HDL-C ratio, can predict the risk of a future atherogenic cardiometabolic event from an early age , 34–36. al study  with 32 Data on the potential effects of ω3-LCPUFAs on anti-inflammatory actions and cardiometabolic factors in children with ALL are scarce. Nevertheless, there is evidence that ω3-LCPUFAs, reduce inflammatory markers in adult oncology patients –41, as eBased on these results, we should discuss the strengths and limitations of our work. A strength of the study was the use of a prospective cohort, which improved the accuracy of data collection. Another added benefit was the use of controls. In addition, supplementation at a rate of 0.100 g/kg/d   during eAlthough ω3-LCPUFAs have a positive effect on cancer patients, in ALL pediatric patients there is no sufficient data for recommendations. As we stated above, available studies from randomized placebo-controlled trials the supplements evaluated have varied in dose, source, time of intervention and kind of supplements used in these of ω3-LCPUFAs , 24, 44.Dyslipidemia and inflammation have some common pathological links, such as obesity and cancer. Recent literature mentions that lipids have a fundamental role in the activation of inflammatory pathways, thus increasing the production of inflammatory cytokines , which can promote the interruption of lipid metabolism, especially the reverse transport of cholesterol; this is related to a decrease in HDL-C, which could stimulate compensatory changes, such as the synthesis and accumulation of VLDL-C and hypertriglyceridemia, and ultimately increase cardiometabolic risk . On the These findings support the use of omega-3 fatty acids to reduce some adverse cardiometabolic and inflammatory risk factors in children with ALL. Our findings show that an ω3-LCPUFA intervention is feasible, and the results suggest that ω3-LCPUFA supplementation in these patients could help prevent, delay, and/or mitigate the development of dyslipidemia and cardiometabolic conditions that can have a negative impact on them. It is unknown if the effect of omega-3 may last longer than 3 months. We consider that large-scale trials are needed in children with cancer to confirm these results.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.This study was approved by the Research and Ethics Committee of the Pediatric Hospital at the Mexican Social Security Institute (2009-785-107). We obtained written informed consent from parents and informed assent from children. Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin.LB-C, designed the study, analyzed data, obtained funding, and prepared the manuscript; SA-M, EJ-A, JM-T and FM-B, contributed to the analysis and interpretation of data and helped prepare the manuscript; ML-A, JM-A performed the analysis and interpretation of data, and helped prepare the manuscript; SD-P, BG, JD-P, AA-B, human resource management, contributed to the analysis and interpretation of data, S-LK, BB-M, AJ-M, ZH-P, EJ-H, LE-H, NN-V, RP-C contributed to the acquisition of data. All authors contributed to the article and approved the submitted version.